AG490在膀胱癌中抗癌效应及其机制.docVIP

AG490在膀胱癌中抗癌效应及其机制 　 作者：姚林方，叶章群，陈志强，刘冠琳，孔德波，杨为民 【摘要】 目的 观察JAK2激酶抑制剂AG490对膀胱癌细胞增殖和凋亡的影响，并探讨其抗癌机制。方法 应用不同剂量AG490处理膀胱癌细胞系BIU87，台盼蓝排斥试验检测细胞活力，噻唑蓝比色试验检测细胞增殖，克隆形成试验进一步检测膀胱癌细胞系干细胞对药物的敏感性，Hoechst33258/PI荧光双染检测细胞凋亡特征，流式细胞仪检测细胞周期和凋亡，Western blot检测pJAK2、STAT3、pSTAT3、Cyclin D1、bclxL蛋白表达水平；并建立膀胱癌荷瘤模型，观察AG490体内抗肿瘤作用。结果 AG490明显抑制膀胱癌细胞增殖和干细胞克隆形成，使细胞周期阻滞，促进膀胱癌细胞凋亡；并可使pJAK2、STAT3、pSTAT3、Cyclin D1、bclxL蛋白表达水平明显下降。裸鼠移植瘤在AG490的作用下明显缩小。结论 AG490通过阻断STAT3信号通路，抑制膀胱癌细胞的增殖，促进其调亡。 【关键词】 膀胱癌；AG490；信号通路；抗癌效应 　　The Mechanism of Anticancer Effects of Janus Kinase Inhibitor AG490 in Human Bladder Cancer 　　　Abstract：Objective To investigate the anticancer effects of Janus Kinaseselective inhibitor AG490 in human bladder cancer.Methods The bladder cancer cell line BIU87 was treated with AG490 in different doses. The cell vitality and proliferation were detected by Trypan Blue staining rejection, cell counting and MTT assay. Drug sensitivity of bladder cancer stem cells to AG490 was detected by clone formation counting assay. Fluorescence dyestuff Hoechst33258 and PI doublestaining assay was used to investigate the cell apoptosis characteristics. Flow cytometry was applied to analyze the cell cycle and apoptosis. The expressions of phosphorylationspecific JAK2(pJAK2), STAT3, phosphorylationspecific STAT3(pSTAT3), Cyclin D1 and bclxL were measured by western blot.Results AG490 could remarkably inhibit the proliferation of bladder cancer cells and the formation of the tumor stem cell clones, block the cell cycle, facilitate the apoptosis of bladder cancer cells, and result in less expression of pJAK2, STAT3, pSTAT3, Cyclin D1 and bclxL.Conclusion AG490 showed strong abilities of inhibiting the proliferation of bladder cancer cells and inducing their apoptosis through blocking the STAT3 signaling pathway. 　　Key words：Bladder cancer；AG490；Signaling pathway；Anticancer effects 　　0 引言 膀胱癌是我国泌尿系最常见的恶性肿瘤，发病率居泌尿系肿瘤首位，至今仍无特异的抗癌药物。信号转导和转录激活因子3(Signal transducer and activator of transcription 3